Extracellular Vesicles

Extracellular Vesicles

Extracellular vesicles (EVs) are small membranous structures that are released from a variety of cell types. These can be found in various bodily fluids, including blood, urine, and saliva. These are known to play critical roles in intercellular communication and can contain a variety of bioactive molecules, including proteins, lipids, and nucleic acids. Given their importance in both normal physiological processes and disease states, interest has increased  in the analysis of EVs.

The first experiments in which EVs were studied began in the 80’s. The interest for these vesicles has been growing ever since, with an exponential development of the field over the past two decades.

EV analysis can involve a wide range of techniques that are essential for characterizing the size, concentration, and cargo of EVs, as well as for identifying the specific cell types that release it. Additionally, EV analysis can provide insights into the mechanisms by which EVs are taken up by recipient cells and the biological effects these induce.

What are EVs?

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What are EVs?

Different categories of EVs

Extracellular vesicles (EVs) can be broadly classified into three categories based on their biogenesis and size.

Exosomes: These are small EVs (30-150 nm) that are derived from the endosomal pathway. They are formed by the inward budding of the endosomal membrane and the subsequent fusion of multivesicular bodies (MVBs) with the plasma membrane. Exosomes carry various biomolecules, including proteins, lipids, and nucleic acids, and are involved in intercellular communication and cargo delivery.

Microvesicles: These are larger EVs (100-1000 nm) that are formed by the outward budding of the plasma membrane. Microvesicles carry a variety of bioactive molecules, including growth factors, cytokines, and signaling lipids. They are involved in intercellular communication, immune regulation, and blood clotting.

Apoptotic bodies: These are large EVs (1-5 µm) that are released by cells undergoing programmed cell death (apoptosis). Apoptotic bodies carry a diverse range of molecules, including chromatin, enzymes, and cellular organelles. These are involved in the clearance of apoptotic cells by phagocytes and the maintenance of tissue homeostasis.

We can consider small EVs (<200 nm) and medium/large EVs (<200 nm).

What are EVs used for?

They are involved in various physiological and pathological processes, and researchers have extensively studied its potential applications in medicine and biotechnology. Extracellular vesicles have a wide range of uses in various, and their therapeutic potential is currently being explored in many preclinical and clinical studies.

Intercellular communication: EVs play an important role in intercellular communication by transferring biological molecules, such as proteins, lipids, and nucleic acids, between cells. This process can influence various physiological and pathological processes, including immune response, tissue regeneration, and cancer progression.

Diagnostic markers: EVs can serve as diagnostic markers for various diseases, as it contains biomolecules that reflect the physiological state of the parent cell. For example, circulating EVs have been shown to contain biomarkers for cancer and neurodegenerative diseases, and their analysis can aid in disease diagnosis and prognosis.

Drug delivery: EVs can be engineered to deliver therapeutic molecules, such as drugs or nucleic acids, to target cells. These offer several advantages over traditional drug delivery systems, including high biocompatibility, low immunogenicity, and the ability to cross biological barriers.

Tissue engineering: EVs can promote tissue regeneration and repair by modulating cellular behavior and signaling pathways. These have been used in various tissue engineering applications, including bone regeneration, cartilage repair, and wound healing.

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HORIBA Solutions

HORIBA Scientific instrumentation can help in the characterization of extracellular vesicles (EVs) in several ways:

Size and concentration measurements – ViewSizer3000

HORIBA's nanoparticle tracking analysis (NTA) systems can accurately measure the size and concentration of EVs. NTA is a powerful tool for EV analysis, as it allows for the simultaneous measurement of particle size and concentration in real-time. Additionally, the use of fluorescent labeling with NTA enables the discrimination of different EV subpopulations based on its biomolecular content.

  • Accurate size distribution of polydisperse samples
  • No calibration standards required
  • Particle concentration measurement
  • Wide dynamic size range (10 nm – 5 μm)
  • No cross contamination
  • Visualize particles and processes
  • Individual particle method

Zeta potential and size measurements – nanoPartica SZ-100V2

The zeta potential of EVs can provide valuable information about their surface charge and stability. HORIBA's zeta potential analyzers can measure the zeta potential of EVs, allowing researchers to understand how EVs interact with other biological structures. nanoPartica SZ-100V2 can also provide information about the size of extracellular vesicles.

  • EV size distribution
  • EV mean size determination
  • Zeta potential determination
  • Ensure colloidal stability of EVs in suspension

Chemical identification of EVs – LabRAM Soleil

Raman spectroscopy is a powerful analytical technique that can be used to identify the chemical composition of EVs. This technique is ideal for EV characterization, as it can provide high-resolution spectra with minimal sample preparation.

Raman spectroscopy can be combined with the AFM technique (TERS) for a more efficient analysis and to get nanoscale chemical and structural information and topography distribution, making the AFM-Raman platform a powerful tool.

  • Mapping of chemical composition
  • Vesicle visualization
  • Quality control
  • Purity control
  • Mechanical properties

A-TEEM for molecular fingerprinting of EVs - Aqualog

A-TEEM fluorescence method quickly fingerprints molecules, differentiates exosome types, detects contaminants, and studies the biodistribution of extracellular vesicles. It is a non-destructive and label-free tool, insensitive to many common excipients while being hypersensitive to proteins and consequently to EVs. A-TEEM profiles provide better insights into protein signatures in complex matrices. This makes A-TEEM a valuable tool for QA/QC and process analysis.

  • Unique molecular fingerprint of Evs
  • EVs phenotyping
  • Sample-to-sample variation
  • Aggregation evaluation
  • Contaminant detection
  • Molecular binding
  • Solvant effect visualization

Biological interaction – OpenPlex

The main interest of Surface Plasmon Resonance imaging is its ability to provide information about the presence of specific surface markers. This information can be used to distinguish different subpopulations of EVs, which may have different biological functions.

  • Biomolecular interaction
  • Kinetics profile / Affinity determination
  • Label-free
  • Measurement in crude media

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Application Notes and Webinars

Featured Application Notes

Read the Readout Article

Providing Solutions for the Life Science Field Using Spectroscopic Analyzers (Readout No. E55)

READOUT is a technical journal issued by HORIBA.

Featured Webinars

Exosomes: exploiting the diagnostic and therapeutic potential of Nature’s biological nanoparticles

Research on exosomes and other forms of extracellular vesicles (EVs) have rapidly expanded over the last two decades. These lipid-enclosed, nanoscale messengers are released from cells packed with diverse cargo and can travel long distances to modify the function of target cells. Found in abundant quantities in biological fluids like blood, there is great clinical interest in using EVs as diagnostic markers or altering their properties for therapeutic delivery. Tune in to find out more about what exosomes are, how researchers study these, and what challenges remain. This talk will highlight multi-laser nanoparticle tracking analysis (NTA) with the ViewSizer 3000 and what it offers in exosome research

Extracellular Vesicle Secretion: Tissue-Specificity and the Impact of Health and Disease

Professor Dan Lark, director of the “Extracellular Regulation of Metabolism” laboratory at Colorado State University in the Department of Health & Exercise Science will be giving a talk through the WebEVTalk series focusing on their findings in EV secretion between skeletal muscle and white adipose tissue. This study uses transgenic fluorescent reporter mice and various analytical techniques including the multi-laser Nanoparticle Tracking Analysis (m NTA) to collectively demonstrate improved understanding of EV secretion by different tissues and their biodistribution. Link to paper: click here.

WebEVTalk series, founded by Dr. Carolina Soekmadji, is a regular talk series for EV enthusiasts around the globe. Dr. Soekmadji is a Principal Investigator at QIMR Berghofer Medical Research Institute and an active editorial board member for Journal of Extracellular Vesicles @ISEV_JEV

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Raman


SERS

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Extracellular Vesicles Brochure

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